銀葉病菌のペクチン酸リア-ゼIとIIの精製と性質 Purification and Characterization of Pectate Lyase I and II from Fungus Stereum purpureum

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Author(s)

    • 宮入 一夫 MIYAIRI Kazuo
    • 弘前大学農学生命科学部応用生命工学科 Department of Biochemistry and Biotechnology, Faculty of Agriculture and Life Science, Hirosaki University
    • 西田 佳織 NISHIDA Kaori
    • 弘前大学農学生命科学部応用生命工学科 Department of Biochemistry and Biotechnology, Faculty of Agriculture and Life Science, Hirosaki University
    • 宝栄 優 [他] TAKARAE Masaru
    • 弘前大学農学生命科学部応用生命工学科 Department of Biochemistry and Biotechnology, Faculty of Agriculture and Life Science, Hirosaki University
    • 鹿内 由美子 SHIKANAI Yumiko
    • 弘前大学農学生命科学部応用生命工学科 Department of Biochemistry and Biotechnology, Faculty of Agriculture and Life Science, Hirosaki University
    • 奥野 智旦 OKUNO Toshikatsu
    • 弘前大学農学生命科学部応用生命工学科 Department of Biochemistry and Biotechnology, Faculty of Agriculture and Life Science, Hirosaki University

Abstract

リンゴ銀葉菌の培養口液からペクチン酸リアーゼIとII(PL I,II)をSDS-PAGE的に均一成分として単離した.これらの分子質量は48kDaで等しく,Endo H処理によりともに38kDaに減少した.また,等電点はそれぞれ4.15と4.10であった.N末端アミノ酸20残基は共通であり,アミノ酸組成もきわめて似ていた.これらは至適pHが95,熱安定性は50℃ まで,pH安定性はpH6.0-7.0とほぼ同じであり,0.2mMCa<SUP>2+</SUP>存在下で最大活性を示した.反応生成物のHPLC分析で,反応初期からオリゴガラクツロン酸を生成したことからエンド型と推定されたが,E . carotovora PLによる生成物パターンとは異なっていた.また,反応最終産物は不飽和のジとトリガラクツロン酸であったが,その比率はE.carotovora PLの場合とは異なっていた.

Two pectate lyases (PLs I and II) were isolated from the culture filtrate of Stereum purpureum, the causative fungus of apple silver-leaf disease. The lyases were isolated using DE52 column chromatography, and their molecular masses were estimated to be 48 kDa by SDS-PAGE. After treatment with endo-β-N-acetylglucosaminidase (Endo H), their molecular masses decreased equally to 38 kDa, indicating that these PLs are glycoproteins. The pI values for PLs I and II were determined to be 4.15 and 4.10, respectively. The amino acid compositions of PLs I and II were very similar, and furthermore the amino acid sequences of the N-terminal 20 residues of the two enzymes were identical. Their optimum pH values, and thermal and pH stabilities were 9.5, up to 50°C, and pH 6-7, respectively. Both PLs I and II required 0.2 mM Ca<SUP>2+</SUP> ion for maximum activity. The enzymes were assigned as endo-types because of the detection of oligo-GalUAs as products from the initial reaction. However they mainly formed products less than octa-GaIUA in degree of polymerization. Erwinia carotovora PL mainly formed products more than octa-GalUA. Therefore they were different in their initial reaction products. As in the case of the E. carotovora PL, the final products of S. purpureum PL were 4, 5-unsaturated di- and tri-GalUA.

Journal

  • Journal of Applied Glycoscience

    Journal of Applied Glycoscience 49(2), 99-106, 2002-04-01

    The Japanese Society of Applied Glycoscience

References:  28

Codes

  • NII Article ID (NAID)
    10008253442
  • NII NACSIS-CAT ID (NCID)
    AN10453916
  • Text Lang
    ENG
  • Article Type
    ART
  • ISSN
    13403494
  • NDL Article ID
    6142235
  • NDL Source Classification
    ZP24(科学技術--化学・化学工業--糖・澱粉)
  • NDL Call No.
    Z17-15
  • Data Source
    CJP  NDL  J-STAGE  JASI 
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